title: rmpM genosensor for detection of human brain bacterial meningitis in cerebrospinal fluid.
abstract: Human brain bacterial meningitis is a life-threatening disease caused mainly by Neisseria meningitidis, lead to damage of the outer membrane covering (meninges) of brain or even death. The usual methods of diagnosis are either time-consuming or have some limitations. The specific rmpM (reduction-modifiable protein M) virulent gene based genosensor is more sensitive, specific, and can detect N. meningitidis directly from the patient cerebrospinal fluid in 30 min including 1-min response time. 5'-Thiol-labeled single-stranded DNA (ssDNA) probe was immobilized onto screen-printed gold electrode (SPGE) and hybridized with denatured (95 °C) single-stranded genomic DNA (ssG-DNA) for 10 min at 25 °C. The electrochemical response was measured by cyclic voltammetry, differential pulse voltammetry (DPV) and electrochemical impedance using redox indicators. The sensitivity of the genosensor was 9.5087 (μA/cm(2))/ng with DPV and limit of detection was 3 ng/6 μL ssG-DNA. The immobilization of the ssDNA probe and hybridization with ssG-DNA from N. meningitidis was characterized by atomic force microscopy and Fourier transform infrared spectroscopy. The rmpM genosensor was stable for 6 months at 4 °C with 10 % loss in initial DPV current. The advantage of rmpM genosensor is to detect bacterial meningitis simultaneously in multiple patients using SPGE array during an outbreak of the disease.
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, 110007, India.
authors.forename: Sandip Kumar
authors.initials: SK
authors.lastname: Dash
authors.forename: Minakshi
authors.initials: M
authors.lastname: Sharma
authors.forename: Shashi
authors.initials: S
authors.lastname: Khare
authors.forename: Ashok
authors.initials: A
authors.lastname: Kumar
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D015374:Biosensing Techniques / Q000379:methods*; D001921:Brain / Q000382:microbiology*; D006801:Humans; D016920:Meningitis, Bacterial / Q000134:cerebrospinal fluid* / Q000382:microbiology*; D009345:Neisseria meningitidis / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology
chemical_list: D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C499670:RmpM protein, Neisseria